基于网络药理学、分子对接及动物试验研究虎杖防治雏鹅痛风的作用机制  

Study on the mechanism of action of Polygonum cuspidatum against gout in goslings based on network pharmacology,molecular docking and animal testing

作　　者：朱道仙 吴植[1] 郝福星 刘静[2] 卢劲晔[2] ZHU Daoxian;WU Zhi;HAO Fuxing;LIU Jing;LU Jinye(Department of Veterinary Medicine,Jiangsu Agri-animal Husbandry Vocational College,Taizhou 225300,China;Department of Pet Science and Technology,Jiangsu Agri-animal Husbandry Vocational College,Taizhou 225300,China)

机构地区：[1]江苏农牧科技职业学院动物医学院,江苏泰州225300 [2]江苏农牧科技职业学院宠物科技学院,江苏泰州225300

出　　处：《黑龙江畜牧兽医》2024年第13期91-97,113,114,共9页Heilongjiang Animal Science And veterinary Medicine

基　　金：江苏省高校教师青蓝工程学科带头人项目(2021);江苏农牧科技职业学院校级科研课题(NSF2021ZR17)。

摘　　要：为了探讨虎杖防治痛风的潜在作用机制,试验采用网络药理学方法,通过中药系统药理数据库和分析平台(TCMSP)筛选出虎杖的活性成分、作用靶点,通过GeneCards数据库获取痛风的相关靶点;取虎杖与痛风的相交靶点,用Cytoscape 3.8.2软件构建“中药-活性成分-靶点-疾病”网络;用String数据库构建蛋白质-蛋白质相互作用(PPI)关系图,找出关键靶点;对靶点进行KEGG信号通路富集分析,构建“活性成分-靶点-通路”网络;选取关键靶点和主要活性成分,用Autodock 1.5.6软件进行分子对接;建立雏鹅痛风模型,用qRT-PCR检测相关靶蛋白的mRNA表达,验证虎杖的网络药理学分析结果。结果表明:共得到虎杖活性成分10个,作用靶点183个,痛风相关靶点1557个,虎杖防治痛风潜在靶点71个。PPI分析发现关键靶点8个,分别为RELA、MAPK1、Akt1、NFKBIA、TNF、IL-6、MAPK14及HSP90AA1等,主要富集于TNF、NF-κB、Toll样受体、NOD样受体和HIF-1等5条信号通路中。RELA、MAPK1、NFKBIA及TNF等靶点的结合能力较强,能与虎杖中槲皮素、木犀草素稳定结合。虎杖能显著提高痛风雏鹅体重和治愈率(P<0.05),显著降低血清中尿酸和肌酐含量及TNF-α、IL-6、IL-1β等炎症因子水平及肾脏中RELA、TNF及MAPK1等mRNA相对表达量(P<0.05)。说明虎杖可能通过槲皮素、山奈酚等活性成分调控痛风过程中RELA、MAPK1、NFKBIA及TNF等靶点,从而降低炎症因子活性,减轻炎症反应,恢复肾脏功能。In order to explore the potential mechanism of Polygonum cuspidatum on gout prevention and treatment,a network pharmacology approach was adopted to screen out the active ingredients and targets of Polygonum cuspidatum through Traditional Chinese Medicine Systematic Pharmacology(TCMSP)database and analytical platform.The Gene Cards database was used to obtain the relevant targets of gout;the intersecting targets of Polygonum cuspidatum and gout were analyzed,and the"Traditional Chinese Medicine-Active Ingredient-Target-Disease"network was constructed using Cytoscape 3.8.2 software;the potential targets of Polygonum cuspidatum were imported into the String database,for the construction of a protein-protein interaction(PPI)diagram to identify the key targets;KEGG signaling pathway enrichment of the targets was analyzed,and a network of"active ingredient-target-pathway"was constructed;the key targets and main active ingredients were selected,and molecular docking was carried out with Autodock 1.5.6 software;finally,a chick gout model was established;the gene expression of relevant target proteins was detected by qRT-PCR to validate the results of the network pharmacological analysis of Polygonum cuspidatum.The results showed that a total of 10 active ingredients,183 action targets,1,557 gout-related targets,and 71 potential targets of Polygonum cuspidatum for the prevention and treatment of gout were obtained.PPI analysis revealed 8 key targets,which were RELA,MAPK1,Akt1,NFKBIA,TNF,IL-6,MAPK14 and HSP90AA1,and were mainly enriched in 5 signaling pathways,including TNF,NF-κB,Toll-like receptor,NOD-like receptor and HIF-1.In molecular docking,the targets with strong binding ability were RELA,MAPK1,NFKBIA and TNF,which could be stably bound to quercetin and luteolin in Polygonum cuspidatum.Compared with the control group,Polygonum cuspidatum significantly increased the body weight and cure rate of gouty chicks(P<0.05),significantly decreased the serum levels of uric acid and creatinine and the levels of inflammatory fa

关 键 词：虎杖 雏鹅 痛风 槲皮素 山奈酚 网络药理学 分子对接 

分 类 号：S856.5[农业科学—临床兽医学] S853.9[农业科学—兽医学]